The Keap1-Nrf2 signaling pathway regulates antioxidant defenses of Ctenopharyngodon idella induced by bacterial infection.

Respiratory burst is a process involving rapid production of reactive oxygen species (ROS) for eliminating invading pathogens. However, excessive ROS production can be fatal to the host organism. The Keap1-Nrf2-ARE (Kelch-like ECH-associated protein 1 [Keap1]; Nuclear factor erythroid-derived 2-like 2 [Nrf2]; Antioxidant responsive element [ARE]) signaling pathway plays an important role in alleviating oxidative stress and preserving cellular homeostasis. However, the role of Keap1 during bacterial infection in fish remains unclear. In this study, we cloned and characterized the Keap1 gene of grass carp (CiKeap1) for the first time. CiKeap1 encodes a 593-amino acid protein of the Keap1b type. The tissue distribution analysis data revealed that the brain contains the highest transcription level of Keap1, followed by the heart and liver. The infection of Aeromonas hydrophila and Staphylococcus aureus obviously modulated the gene transcription and protein expression levels of Keap1, which suggested that the CiKeap1 participates in antibacterial immune responses. Furthermore, in vitro overexpression assays clarified the defensive and regular roles of CiKeap1 in maintaining host redox homeostasis in response to bacterial infection through the Keap1-Nrf2-ARE signaling pathway. In conclusion, the present results provide an expanded perspective on the role of Keap1 in teleost immunology that can guide healthy farming cultivation of grass carp.

Antiproliferative effects of n-butyl-ß-D-fructofuranoside from Kangaisan on Bel-7402 cells.

AIMS: Kangaisan is a powdered compound prescription of Traditional Chinese Medicine which has been used in cancers for many years in Hubei province, China. The purpose of this study was to investigate the antitumor effects of Kangaisan and screen bioactive components. MATERIALS AND METHODS: 3-(4,5-Dimethythiazol-2-yl)-2,5 diphenyl-tetrazolium bromide (MTT) assay, flow cytometry, DNA (Deoxyribonucleic acid) fragmentation assay, Western blot, and real time-polymerase chain reaction were used to investigate the antiproliferation effect of n-butyl-ß-D-fructofuranoside on Bel-7402 cells. STATISTICAL ANALYSIS: All experiments were performed in triplicate and the results were expressed as mean ± standard deviation. Statistical analysis was performed with analysis of variance using Origin 8.0 software. RESULTS: It was illustrated that treatment of Bel-7402 cells with various concentrations of n-butyl-ß-D-fructofuranoside resulted in growth inhibition in both a dose-dependent and time-dependent manner. The arrest of G0/G1 phase was also induced (P < 0.05). The increasing of sub-G1 cell population indicated the apoplectic characteristic (P < 0.05). Furthermore, the emerging of DNA fragmentation and the increase of Bax/Bcl-2 ratio and p53 expression suggested the possible mitochondrial apoptotic pathway (P < 0.05). CONCLUSIONS: The results illustrate that Kangaisan showed anticancer effects and n-butyl-ß-D-fructofuranoside extracted from Kangaisan can suppress Bel-7402 cells via interfering cell cycle and by inducing apoptosis.